U.S. patent application number 12/864001 was filed with the patent office on 2011-01-27 for neuronal cell death inhibitor.
This patent application is currently assigned to LEAD CHEMICAL CO., LTD.. Invention is credited to Yuji Matsuya, Hideo Nemoto, Chihiro Tohda.
Application Number | 20110021625 12/864001 |
Document ID | / |
Family ID | 40900972 |
Filed Date | 2011-01-27 |
United States Patent
Application |
20110021625 |
Kind Code |
A1 |
Nemoto; Hideo ; et
al. |
January 27, 2011 |
NEURONAL CELL DEATH INHIBITOR
Abstract
There is provided a neuronal cell death inhibitor having a high
efficacy. A neuronal cell death inhibitor comprising a compound of
general formula (1) below: ##STR00001## (where R.sup.1 is a
hydrogen atom or an optionally substituted hydroxy group; R.sup.2
is a hydrogen atom; R.sup.3 is a hydrogen atom or an optionally
substituted hydroxy group; R.sup.4 is a hydrogen atom or an
optionally substituted hydroxy group, R.sup.3 together with R.sup.4
is optionally an oxo group or a group of --O--(CH.sub.2).sub.n--O--
(where n is an integer of 2 to 4), or R.sup.2 together with R.sup.4
optionally forms an unsaturated bond between carbon atoms, the
respective carbon atoms being attached to R.sup.2 and R.sup.4;
R.sup.5 is a hydrogen atom or an optionally substituted lower alkyl
group; each of R.sup.6, R.sup.7, and R.sup.8 is the same as or
different from each other and is a hydrogen atom, an optionally
substituted hydroxy group, an optionally substituted alkyl group,
an optionally substituted aryl group, or an optionally substituted
heteroaryl group); or a salt thereof.
Inventors: |
Nemoto; Hideo; (Toyama-shi,
JP) ; Tohda; Chihiro; (Imizu-shi, JP) ;
Matsuya; Yuji; (Imizu-shi, JP) |
Correspondence
Address: |
OLIFF & BERRIDGE, PLC
P.O. BOX 320850
ALEXANDRIA
VA
22320-4850
US
|
Assignee: |
LEAD CHEMICAL CO., LTD.
TOYAMA-SHI, TOYAMA
JP
|
Family ID: |
40900972 |
Appl. No.: |
12/864001 |
Filed: |
January 23, 2009 |
PCT Filed: |
January 23, 2009 |
PCT NO: |
PCT/JP2009/000253 |
371 Date: |
September 23, 2010 |
Current U.S.
Class: |
514/462 ;
549/338 |
Current CPC
Class: |
C07D 407/04 20130101;
C07D 493/10 20130101; A61P 25/16 20180101; A61P 25/00 20180101;
A61P 25/02 20180101; A61P 43/00 20180101; A61P 25/28 20180101 |
Class at
Publication: |
514/462 ;
549/338 |
International
Class: |
A61K 31/357 20060101
A61K031/357; C07D 493/10 20060101 C07D493/10; A61P 25/00 20060101
A61P025/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 23, 2008 |
JP |
2008-012642 |
Claims
1. A neuronal cell death inhibitor comprising: a compound of
general formula (1) below: ##STR00031## (where R.sup.1 is a
hydrogen atom or an optionally substituted hydroxy group; R.sup.2
is a hydrogen atom; R.sup.3 is a hydrogen atom or an optionally
substituted hydroxy group; R.sup.4 is a hydrogen atom or an
optionally substituted hydroxy group, R.sup.3 together with R.sup.4
is optionally an oxo group or a group of --O--(CH.sub.2).sub.n--O--
(where n is an integer of 2 to 4), or R.sup.2 together with R.sup.4
optionally forms an unsaturated bond between carbon atoms, the
respective carbon atoms being attached to R.sup.2 and R.sup.4;
R.sup.5 is a hydrogen atom or an optionally substituted lower alkyl
group; each of R.sup.6, R.sup.7, and R.sup.8 is the same as or
different from each other and is a hydrogen atom, an optionally
substituted hydroxy group, an optionally substituted alkyl group,
an optionally substituted aryl group, or an optionally substituted
heteroaryl group); or a salt thereof.
2. The neuronal cell death inhibitor according to claim 1, wherein
each of R.sup.1 and R.sup.5 is a hydrogen atom.
3. The neuronal cell death inhibitor according to claim 1 wherein
each of R.sup.6, R.sup.7, and R.sup.8 is the same as or different
from each other and is a hydrogen atom; a hydroxy group optionally
substituted with a sulfonyl group, a silyl group, or a lower alkyl
group optionally substituted with a halogen atom; or an aryl group
substituted with a halogen atom.
4. A therapeutic and/or prophylactic agent for a neurological
disorder, comprising: a compound of general formula (1) below:
##STR00032## here R.sup.1 is a hydrogen atom or an optionally
substituted hydroxy group; R.sup.2 is a hydrogen atom; R.sup.3 is a
hydrogen atom or an optionally substituted hydroxy group; R.sup.4
is a hydrogen atom or an optionally substituted hydroxy group,
R.sup.3 together with R.sup.4 is optionally an oxo group or a group
of --O--(CH.sub.2).sub.n--O-- (where n is an integer of 2 to 4), or
R.sup.2 together with R.sup.4 optionally forms an unsaturated bond
between carbon atoms, the respective carbon atoms being attached to
R.sup.2 and R.sup.4; R.sup.5 is a hydrogen atom or an optionally
substituted lower alkyl group; each of R.sup.6, R.sup.7, and
R.sup.8 is the same as or different from each other and is a
hydrogen atom, an optionally substituted hydroxy group, an
optionally substituted alkyl group, an optionally substituted aryl
group, or an optionally substituted heteroaryl group); or a salt
thereof.
5. The therapeutic and/or prophylactic agent for a central nervous
system disease according to claim 4, wherein each of R.sup.1 and
R.sup.5 is a hydrogen atom.
6. The therapeutic and/or prophylactic agent for a central nervous
system disease according to claim 4, wherein each of R.sup.6,
R.sup.7, and R.sup.8 is the same as or different from each other
and is a hydrogen atom; a hydroxy group optionally substituted with
a sulfonyl group, a silyl group, or a lower alkyl group optionally
substituted with a halogen atom; or an aryl group substituted with
a halogen atom.
7-12. (canceled)
13. A method for inhibiting neuronal cell death comprising
administering an effective amount of a compound of general formula
(1) below: ##STR00033## here R.sup.1 is a hydrogen atom or an
optionally substituted hydroxy group; R.sup.2 is a hydrogen atom;
R.sup.3 is a hydrogen atom or an optionally substituted hydroxy
group; R.sup.4 is a hydrogen atom or an optionally substituted
hydroxy group, R.sup.3 together with R.sup.4 is optionally an oxo
group or a group of --O--(CH.sub.2).sub.n--O-- (where n is an
integer of 2 to 4), or R.sup.2 together with R.sup.4 optionally
forms an unsaturated bond between carbon atoms, the respective
carbon atoms being attached to R.sup.2 and R.sup.4; R.sup.5 is a
hydrogen atom or an optionally substituted lower alkyl group; each
of R.sup.6, R.sup.7, and R.sup.8 is the same as or different from
each other and is a hydrogen atom, an optionally substituted
hydroxy group, an optionally substituted alkyl group, an optionally
substituted aryl group, or an optionally substituted heteroaryl
group) or a salt thereof.
14. The method according to claim 13, wherein each of R.sup.1 and
R.sup.5 is a hydrogen atom.
15. The method according to claim 13, wherein each of R.sup.6,
R.sup.7, and R.sup.8 is the same as or different from each other
and is a hydrogen atom; a hydroxy group optionally substituted with
a sulfonyl group, a silyl group, or a lower alkyl group optionally
substituted with a halogen atom; or an aryl group substituted with
a halogen atom.
16. A method for treating and/or preventing a neurological disorder
comprising administering an effective amount of a compound of
general formula (1) below: ##STR00034## (where R.sup.1 is a
hydrogen atom or an optionally substituted hydroxy group; R.sup.2
is a hydrogen atom; R.sup.3 is a hydrogen atom or an optionally
substituted hydroxy group; R.sup.4 is a hydrogen atom or an
optionally substituted hydroxy group, R.sup.3 together with R.sup.4
is optionally an oxo group or a group of --O--(CH.sub.2).sub.n--O--
(where n is an integer of 2 to 4), or R.sup.2 together with R.sup.4
optionally forms an unsaturated bond between carbon atoms, the
respective carbon atoms being attached to R.sup.2 and R.sup.4;
R.sup.5 is a hydrogen atom or an optionally substituted lower alkyl
group; each of R.sup.6, R.sup.7, and R.sup.8 is the same as or
different from each other and is a hydrogen atom, an optionally
substituted hydroxy group, an optionally substituted alkyl group,
an optionally substituted aryl group, or an optionally substituted
heteroaryl group) or a salt thereof.
17. The method according to claim 16, wherein each of R.sup.1 and
R.sup.5 is a hydrogen atom.
18. The method according to claim 16, wherein each of R.sup.6,
R.sup.7, and R.sup.8 is the same as or different from each other
and is a hydrogen atom; a hydroxy group optionally substituted with
a sulfonyl group, a silyl group, or a lower alkyl group optionally
substituted with a halogen atom; or an aryl group substituted with
a halogen atom.
19. A compound of general formula (1a) below: ##STR00035## (where
R.sup.1 is a hydrogen atom or an optionally substituted hydroxy
group; R.sup.2 is a hydrogen atom; R.sup.3 together with R.sup.4 is
a group of --O--(CH.sub.2).sub.n--O-- (where n is an integer of 2
to 4); R.sup.5 is a hydrogen atom or an optionally substituted
lower alkyl group; each of R.sup.6, R.sup.7, and R.sup.8 is the
same as or different from each other and is a hydrogen atom, a
hydroxyl group, a lower alkoxy group, a halogeno lower alkoxy
group, a phenyl lower alkyloxy group, a halogenophenyl lower
alkyloxy group, a lower alkylsulfonyloxy group, a halogeno lower
alkylsulfonyloxy group, an optionally substituted alkyl group, an
optionally substituted aryl group, or an optionally substituted
heteroaryl group, with proviso that cases where each of R.sup.6 and
R.sup.8 is a hydrogen atom and R.sup.7 is a hydrogen atom, an
optionally substituted hydroxyl group, an optionally substituted
alkyl group, an optionally substituted aryl group, or an optionally
substituted heteroaryl group are excluded) or a salt thereof.
20. The compound or a salt thereof according to claim 19, wherein
each of R.sup.1 and R.sup.5 is a hydrogen atom.
21. The compound or a salt thereof according to claim 19, wherein
each of one or more of R.sup.6, R.sup.7 and R.sup.8 is a hydroxyl
group, a lower alkoxy group, a halogeno lower alkoxy group, a
phenyl lower alkyloxy group, a halogenophenyl lower alkyloxy group,
a lower alkylsulfonyloxy group, or a halogeno lower
alkylsulfonyloxy group, and each of the remainder is a hydrogen
atom.
Description
TECHNICAL FIELD
[0001] The present invention relates to a novel neuronal cell death
inhibitor. The present invention specifically relates to a neuronal
cell death inhibitor containing a compound having a tetracyclic
dihydrofuran structure.
BACKGROUND ART
[0002] A neuronal cell includes two neuritis, namely, a dendrite
that receives information from other neuronal cells and an axon
that sends information to other neuronal cells. Neuronal cell death
or neurite atrophy caused by various factors inhibits normal signal
transduction between neuronal cells to cause various diseases
depending on the site of nervous system damage. Specific examples
of the disease caused by a disorder of the central nervous system
(brain and spinal cord) include Parkinson's disease, Alzheimer's
disease, and amytrophic lateral sclerosis, and specific examples of
the disease caused by a disorder of the peripheral nervous system
include multiple neuritis and Guillain-Barre syndrome. In
particular, the central nervous system diseases are progressive,
and effective treatment is not yet available. Neurotrophic
factor-like agents have been studied as therapeutic agents for the
central nervous system diseases. However, such agents mainly have
neuroprotective action and do not clearly show neurite outgrowth
effect in a neurodegenerative environment. In order to treat the
central nervous system diseases, it is required that not only the
neuronal cell death is inhibited but also the neurites of remaining
neuronal cells are extended to normalize the signal transduction
between the neuronal cells. Therefore, what is needed is the
development of a therapeutic agent for central nervous system
diseases having both the neuronal cell death inhibition effect and
the neurite outgrowth effect.
[0003] In contrast, a tetracyclic dihydrofuran compound of general
formula (1) below is known to have antiviral effect (Patent
Document 1 and Non-patent Documents 1 to 3). However, it is
completely unknown how the tetracyclic dihydrofuran compound of
general formula (1) works on the neuronal cells.
Patent Document 1: Japanese Patent Application Publication No.
JP-A-2002-255953. Non-patent Document 1: Bioorg. Med. Chem. 2007,
15, 424-432.
Non-patent Document 2: Heterocycles 2004, 62, 207-211.
[0004] Non-patent Document 3: J. Org. Chem. 2004, 69,
7989-7993.
DISCLOSURE OF THE INVENTION
Problem to be Solved by the Invention
[0005] It is an object of the present invention to provide a novel
neuronal cell death inhibitor and a prophylactic and therapeutic
agent for neurological disorders.
Means for Solving the Problem
[0006] The inventors of the present invention have carried out
intensive studies in order to solve the above problems, and as a
result, have found that the tetracyclic dihydrofuran compound of
general formula (1) below extends the axons and dendrites and
inhibits the neuronal cell death in cerebral cortical neurons of an
Alzheimer's disease rat model treated with amyloid .beta.
(hereinafter called A.beta.). This shows that the neurological
disorders caused by the neuronal cell death or neurite atrophy can
be treated by the tetracyclic dihydrofuran compound of general
formula (1) below. From the above knowledge, the inventors of the
present invention have accomplished the present invention.
[0007] Specifically, the present invention provides a neuronal cell
death inhibitor including a compound of general formula (1)
below:
##STR00002##
[0008] (where R.sup.1 is a hydrogen atom or an optionally
substituted hydroxy group; R.sup.2 is a hydrogen atom; R.sup.3 is a
hydrogen atom or an optionally substituted hydroxy group; R.sup.4
is a hydrogen atom or an optionally substituted hydroxy group,
R.sup.3 together with R.sup.4 is optionally an oxo group or a group
of --O--(CH.sub.2).sub.n--O-- (where n is an integer of 2 to 4), or
R.sup.2 together with R.sup.4 optionally forms an unsaturated bond
between carbon atoms, the respective carbon atoms being attached to
R.sup.2 and R.sup.4; R.sup.5 is a hydrogen atom or an optionally
substituted lower alkyl group; and each of R.sup.6, R.sup.7, and
R.sup.8 is the same as or different from each other and is a
hydrogen atom, an optionally substituted hydroxy group, an
optionally substituted alkyl group, an optionally substituted aryl
group, or an optionally substituted heteroaryl group)
or a salt thereof.
[0009] Furthermore, the present invention provides a therapeutic
and/or prophylactic agent for a neurological disorder, and the
agent includes a compound of general formula (1) or a salt
thereof.
[0010] The present invention also provides a use of a compound of
general formula (1) or a salt thereof for producing a neuronal cell
death inhibitor.
[0011] The present invention also provides a use of a compound of
general formula (1) or a salt thereof for producing a therapeutic
and/or prophylactic agent for a neurological disorder.
[0012] The present invention also provides a method for inhibiting
neuronal cell death characterized by including administering an
effective amount of a compound of general formula (1) or a salt
thereof.
[0013] The present invention also provides a method for treating
and/or preventing a neurological disorder characterized by
including administering an effective amount of a compound of
general formula (1).
[0014] Among the compounds of general formula (1), a compound of
general formula (1a) below is a novel compound. Accordingly, the
present invention provides a compound of general formula (1a)
below:
##STR00003##
[0015] (where R.sup.1 is a hydrogen atom or an optionally
substituted hydroxy group; R.sup.2 is a hydrogen atom; R.sup.3
together with R.sup.4 is a group of --O--(CH.sub.2).sub.n--O--
(where n is an integer of 2 to 4); R.sup.5 is a hydrogen atom or an
optionally substituted lower alkyl group; and each of R.sup.6,
R.sup.7, and R.sup.8 is the same as or different from each other
and is a hydrogen atom, an optionally substituted hydroxy group, an
optionally substituted alkyl group, an optionally substituted aryl
group, or an optionally substituted heteroaryl group, unless each
of R.sup.6 and R.sup.8 is a hydrogen atom and R.sup.7 is a hydrogen
atom, an optionally substituted hydroxy group, an optionally
substituted alkyl group, an optionally substituted aryl group, or
an optionally substituted heteroaryl group)
or a salt thereof.
EFFECTS OF THE INVENTION
[0016] With the medical drug of the present invention, the neurites
can be extended, the neuronal cell death can be remarkably
inhibited, and the neurological disorders such as Alzheimer's
disease can be treated.
BRIEF DESCRIPTION OF THE DRAWINGS
[0017] FIG. 1 shows the neuronal cell death inhibition effect of
Compounds 1 to 5 in rat cerebral cortical neurons treated with
A.beta.(25-35). Numbers in the figure represent the compound
numbers, Cont represents a control group, and Veh represents a
vehicle group. Hereinafter the same applies.
[0018] FIG. 2 shows the neuronal cell death inhibition effect of
Compound 11 in rat cerebral cortical neurons treated with
A.beta.(25-35).
[0019] FIG. 3 shows the dendrite outgrowth effect of Compounds 5
and 11 in rat cerebral cortical neurons treated with
A.beta.(25-35).
[0020] FIG. 4 shows the axon outgrowth effect of Compounds 8 and 11
in rat cerebral cortical neurons treated with A.beta.(25-35).
[0021] FIG. 5 shows the dendrite outgrowth effect of Compounds 5,
11, 21, and 22 in rat cerebral cortical neurons treated with
A.beta.(1-42).
[0022] FIG. 6 shows the axon outgrowth effect of Compounds 5, 11,
21, and 22 in rat cerebral cortical neurons treated with
A.beta.(1-42).
BEST MODES FOR CARRYING OUT THE INVENTION
[0023] In the present specification, the alkyl group means a
C.sub.1-20 alkyl group having a straight chain, a branched chain, a
ring, or a combination thereof, and the lower alkyl group means an
alkyl group having about 1 to 8 carbon atoms, preferably an alkyl
group having about 1 to 6 carbon atoms having a straight chain, a
branched chain, a ring, or a combination thereof. More specific
examples of the lower alkyl group include a methyl group, an ethyl
group, an n-propyl group, an isopropyl group, an n-butyl group, an
isobutyl group, a tert-butyl group, an n-pentyl group, an isopentyl
group, a tert-pentyl group, a hexyl group, a heptyl group, and an
octyl group. Examples of the alkyl group include the lower alkyl
group as well as a decyl group, an undecyl group, a dodecyl group,
a tridecyl group, a tetradecyl group, a pentadecyl group, a
hexadecyl group, a heptadecyl group, an octadecyl group, a
nonadecyl group, and a didecyl group.
[0024] The substituent in "an optionally substituted hydroxy group"
of R.sup.1, R.sup.3, R.sup.4, R.sup.6, R.sup.7, and R.sup.8 in
general formula (1) is not specifically limited, but examples
thereof include an acyl group, a lower alkyl group, a sulfonyl
group, and a silyl group. More specific examples thereof include
acyl groups such as a formyl group, an alkanoyl group, a
phenylcarbonyl group, an alkoxycarbonyl group, and a phenyl lower
alkoxycarbonyl group, lower alkyl groups such as an allyl group, an
alkoxy lower alkyl group, a phenyl lower alkyl group, and a
tetrahydropyranyl group, sulfonyl groups such as a lower
alkylsulfonyl group and an alkoxysulfonyl group, and silyl groups
such as a lower alkylsilyl group and a phenylsilyl group.
[0025] Among the optionally substituted hydroxy groups, more
preferable groups are a hydroxy group, a lower alkoxy group, a
halogen lower alkoxy group, a phenyl lower alkyloxy group, a
halogenophenyl lower alkyloxy group, a lower alkylsulfonyloxy
group, a halogen lower alkylsulfonyloxy group, and a tri-lower
alkylsilyloxy group. Examples of the lower alkoxy group include a
methoxy group, an ethoxy group, a propoxy group, and an
isopropyloxy group. Examples of the halogeno lower alkoxy group
include a trifluoromethoxy group and a trichloromethoxy group.
Examples of the phenyl lower alkyloxy group include a benzyloxy
group and a phenethyloxy group. Examples of the halogenophenyl
lower alkyloxy group include a fluorophenylmethoxy group, a
difluorophenylmethoxy group, and a trifluorophenylmethoxy group.
Examples of the lower alkylsulfonyloxy group include a
methanesulfonyloxy group and an ethanesulfonyloxy group. Examples
of the halogeno lower alkylsulfonyloxy group include a
trifluoromethanesulfonyloxy group. Furthermore, examples of the
tri(lower alkyl)silyloxy group include a trimethylsilyloxy group
and a triisopropylsilyloxy group.
[0026] In "an optionally substituted lower alkyl group" of R.sup.5
and "an optionally substituted alkyl group", "an optionally
substituted aryl group", and "an optionally substituted heteroaryl
group" of each of R.sup.6, R.sup.7, and R.sup.8 in general formula
(1), the substituent, the substitution site, and the number of the
substituents are not specifically limited. Examples of the
substituent include a hydroxy group, an alkoxy group (the alkyl
moiety is the same as that in the alkyl group above), a carboxy
group, a halogen atom (any of a fluorine atom, a chlorine atom, a
bromine atom, or an iodine atom), an optionally substituted amino
group (examples of the substituent include an alkyl group and an
acyl group), an optionally substituted aryl group (examples of the
substituent include an alkyl group, an acyl group, and a halogen
atom), and an optionally substituted heteroaryl group (examples of
the substituent include an alkyl group, an acyl group, and a
halogen atom).
[0027] Specific examples of the optionally substituted lower alkyl
group include a lower alkyl group, a hydroxy lower alkyl group, and
a halogen lower alkyl group. Specific examples of the optionally
substituted aryl group include a phenyl group and a halogenophenyl
group. Specific examples of the optionally substituted heteroaryl
group include a pyridyl group and a furyl group.
[0028] In the compound of general formula (1), each of R.sup.1 and
R.sup.5 is preferably a hydrogen atom. In the compound of general
formula (1), it is preferable that R.sup.2 is a hydrogen atom and
that R.sup.3 together with R.sup.4 is a group of
--O--(CH.sub.2).sub.n--O-- (where n is an integer of 2 to 4), and
it is more preferable that R.sup.2 is a hydrogen atom and that
R.sup.3 together with R.sup.4 is the group of
--O--(CH.sub.2).sub.2--O--.
[0029] In the compound of general formula (1), it is preferable
that each of R.sup.6, R.sup.7, and R.sup.8 is the same as or
different from each other and is a hydrogen atom; an optionally
substituted hydroxy group; or an optionally substituted aryl group.
Among them, it is more preferable that each of R.sup.6, R.sup.7,
and R.sup.8 is the same as or different from each other and is a
hydrogen atom; a hydroxy group optionally substituted with a
sulfonyl group, a silyl group, or a lower alkyl group optionally
substituted with a halogen atom; or an aryl group substituted with
a halogen atom.
[0030] It is more preferable that each of R.sup.6, R.sup.7, and
R.sup.8 is the same as or different from each other and is a
hydrogen atom; a hydroxy group, a lower alkoxy group, a halogen
lower alkoxy group, or a tri(lower alkyl)silyloxy group, it is even
more preferable that each of one or more of R.sup.6, R.sup.7, and
R.sup.8 is a hydroxy group, a lower alkoxy group, a halogen lower
alkoxy group, or a tri-lower alkylsilyloxy group and that each of
the remainder is a hydrogen atom, and it is yet even more
preferable that each of one or more of R.sup.6, R.sup.7, and
R.sup.8 is a hydroxy group or a lower alkoxy group and that each of
the remainder is a hydrogen atom.
[0031] It is yet even more preferable that R.sup.6 is a hydrogen
atom, that each of one or more of R.sup.7 and R.sup.8 is a hydroxy
group or a lower alkoxy group, and that the remain is a hydrogen
atom, and it is most preferable that R.sup.6 is a hydrogen atom,
that R.sup.7 is a hydrogen atom, a hydroxy group, or a lower alkoxy
group, and that R.sup.8 is a hydroxy group or a lower alkoxy
group.
[0032] The compound of general formula (1) can be produced, for
example, by the methods described in Japanese Patent Application
Publication No. JP-A-2002-255953 and Bioorg. Med. Chem. 2007, 15,
424-432.
[0033] The compound of general formula (1) in which R.sup.8 is a
hydroxy group can be produced according to Process A below.
##STR00004##
[0034] (where R.sup.9 is a protective group for the hydroxy group;
and each of R.sup.1 to R.sup.7 is the same as that in the
above).
[0035] In Process A, a protective group of R.sup.9 for the hydroxy
group in the compound of general formula (2) is deprotected to
produce a compound of general formula (3).
[0036] A conventional protective group may be used for the
protective group of R.sup.9 for the hydroxy group, and examples
thereof include a benzyl group, an acetyl group, and an alkylsilyl
group (such as a trimethylsilyl group, a tert-butyldimethylsilyl
group, a triisopropylsilyl group, and a tert-butyldiphenylsilyl
group).
[0037] The deprotection may be carried out in usual ways such as
reduction, hydrolysis, acid treatment, and fluoride treatment
depending on the type of the protective group to be removed.
[0038] In the case of the reduction, for example, the treatment may
be carried out in an appropriate solvent (such as methanol and
ethanol) in a hydrogen atmosphere using a catalyst (such as
palladium-carbon and platinum).
[0039] In the case of the hydrolysis, the treatment may be carried
out in an appropriate solvent (such as tetrahydrofuran, dioxane,
methanol, ethanol, and water) with a base (such as sodium
hydroxide, potassium hydroxide, lithium hydroxide, sodium
methoxide, and sodium ethoxide).
[0040] In the case of the acid treatment, the treatment may be
carried out in an appropriate solvent (such as methanol and
ethanol) with an acid (such as hydrochloric acid, p-toluenesulfonic
acid, methanesulfonic acid, and trifluoroacetic acid).
[0041] In the case of the fluoride treatment, the treatment may be
carried out in an appropriate solvent (such as methanol, ethanol,
acetonitrile, and tetrahydrofuran) with a fluoride (such as
hydrogen fluoride, hydrogen fluoride-pyridine, and
tetrabutylammonium fluoride).
[0042] The reaction temperature varies depending on the
deprotection method, the reaction reagent, and the like, but the
reaction is usually carried out at -50.degree. C. to 100.degree. C.
and preferably at 0.degree. C. to 50.degree. C. The reaction time
varies depending on the reaction temperature, the deprotection
method, the reaction reagent, and the like, but the reaction is
usually carried out for 30 minutes to 48 hours and preferably for 1
hour to 24 hours.
[0043] The compound of general formula (2) as the starting material
in Process A can be produced by the method described in Bioorg.
Med. Chem. 2007, 15, 424-432.
[0044] The compound of general formula (1) in which R.sup.7 is an
optionally substituted aryl group can be produced according to
Process B below.
##STR00005##
[0045] (where R.sup.10 is a halogen atom or a
trifluoromethanesulfonyloxy group; R.sup.11 is an optionally
substituted aryl group; and each of R.sup.1 to R.sup.6 and R.sup.8
is the same as that in the above).
[0046] In Process B, a compound of general formula (4) and boronic
acid compounds of general formula (5) are subjected to Suzuki
coupling reaction in the presence of a base and a palladium
catalyst to produce a compound of general formula (6).
[0047] The solvent used in Process B is not specifically limited as
long as the solvent does not inhibit the coupling reaction.
Examples thereof include ethers such as diethyl ether, diisopropyl
ether, tetrahydrofuran, dioxane, and 1,2-dimethoxyethane, alcohols
such as methanol, ethanol, and propanol, hydrocarbons such as
benzene, toluene, xylene, carbon disulfide, cyclohexane, and
hexane, amides such as formamide, N,N-dimethylformamide, and
N,N-dimethylacetamide, halogenated hydrocarbons such as
dichloromethane, chloroform, carbon tetrachloride, and
1,2-dichloroethane, nitriles such as acetonitrile, propionitrile,
and isobutyronitrile, sulfoxides such as dimethylsulfoxide, water,
and a mixed solvent thereof.
[0048] Examples of the palladium catalyst used in Process B include
tetrakis(triphenylphosphine)palladium, palladium acetate, palladium
chloride, 1,1-bis(diphenylphosphino)ferrocenedichloropalladium, and
dichlorobis(triphenylphosphine)palladium.
[0049] Examples of the base used in Process B include basic salts
such as sodium carbonate, potassium carbonate, cesium carbonate,
and sodium hydrogen carbonate, aromatic amines such as pyridine and
lutidine, tertiary amines such as triethylamine, tripropylamine,
tributylamine, cyclohexyldimethylamine, 4-dimethylaminopyridine,
N,N-dimethylaniline, N-methylpiperidine, N-methylpyrrolidine, and
N-methylmorpholine.
[0050] The reaction temperature varies depending on the solvent,
the catalyst, the base, and the like, but the reaction is usually
carried out at 0.degree. C. to 250.degree. C. and preferably at
50.degree. C. to 150.degree. C. The reaction time varies depending
on the reaction temperature, the solvent, the base, and the like,
but the reaction is usually carried out for 5 minutes to 48 hours
and preferably for 30 minutes to 24 hours.
[0051] The compound of general formula (4) as the starting material
in Process B can be produced by a conventionally known production
method, for example, by triflation of the compound of general
formula (1) in which R.sup.7 is a hydroxy group described in J.
Org. Chem. 2004, 69, 7989-7993 using trifluoromethanesulfonic
anhydride and the like.
[0052] The compound of general formula (1) produced by each of the
above processes can be purified and isolated by a conventionally
known separation and purification technique such as concentration,
solvent extraction, filtration, recrystallization, and various
chromatographic methods.
[0053] The compound of general formula (1) may form a salt. The
salt type is not specifically limited, but generally, when a basic
group such as an amino group is included in the molecule, an acid
addition salt is formed. In contrast, when an acidic group such as
a carboxy group and a phenolic hydroxy group is included in the
molecule, a base addition salt is formed. Examples of the acid
addition salt include a mineral acid salt such as a hydrochloride,
a sulfate, and a nitrate and an organic acid salt such as a
p-toluenesulfonate, a methanesulfonate, an oxalate, a fumarate, a
maleate, and an acetate. Furthermore, examples of the base addition
salt include metal salts such as a sodium salt, a potassium salt, a
calcium salt, and a magnesium salt, an ammonium salt, and organic
amine salts such as a methylamine salt and an ethylamine salt. An
amino acid addition salt such as a glycine salt may also be
formed.
[0054] The compound of general formula (1) has three asymmetric
carbons in the ring and may have further one or more asymmetric
carbons depending on the type of a substituent. Each of the
asymmetric carbons may have any of an R or S configuration. In the
range of the compound of general formula (1), stereoisomers such as
optically active compounds and diastereoisomers based on the
asymmetric carbons, any mixture of the stereoisomers, and racemates
are included. Furthermore, the range of the compound of general
formula (1) includes the compound of general formula (1) and a salt
thereof as well as any hydrates and solvates thereof.
[0055] Among the compounds of general formula (1), the compounds of
(1) to (16) described below are exemplified as specifically
preferred compound. [0056] (1)
5a,10c-dihydro-1H,6H-5-oxa-8-(trifluoromethoxy)acephenanthrylene-10b-carb-
onitrile (hereinafter called Compound 1) [0057] (2)
8-trifluoromethoxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthryl-
ene-10b-carbonitrile ethylene ketal (hereinafter called Compound 2)
[0058] (3)
8-triisopropylsilyloxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephe-
nanthrylene-10b-carbonitrile ethylene ketal (hereinafter called
Compound 3) [0059] (4)
8-methoxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-c-
arbonitrile ethylene ketal (hereinafter called Compound 4) [0060]
(5)
8-hydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-c-
arbonitrile ethylene ketal (hereinafter called Compound 5) [0061]
(6)
8-trifluoromethanesulfonyloxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-ace-
phenanthrylene-10b-carbonitrile ethylene ketal (hereinafter called
Compound 6) [0062] (7)
8-benzyloxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-
-carbonitrile ethylene ketal (hereinafter called Compound 7) [0063]
(8)
8-(3,5-difluorobenzyloxy)-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephen-
anthrylene-10b-carbonitrile ethylene ketal (hereinafter called
Compound 8) [0064] (9)
8-phenyl-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-ca-
rbonitrile ethylene ketal (hereinafter called Compound 9) [0065]
(10)
8-(2,4,6-trifluorophenyl)-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephen-
anthrylene-10b-carbonitrile ethylene ketal (hereinafter called
Compound 10) [0066] (11)
9-hydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-c-
arbonitrile ethylene ketal (hereinafter called Compound 11). [0067]
(12)
7-hydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-c-
arbonitrile ethylene ketal (hereinafter called Compound 12) [0068]
(13)
8,9-dihydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-1-
0b-carbonitrile ethylene ketal (hereinafter called Compound 13)
[0069] (14)
7,9-dihydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthryl-
ene-10b-carbonitrile ethylene ketal (hereinafter called Compound
14) [0070] (15)
7,8,9-trihydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylen-
e-10b-carbonitrile ethylene ketal (hereinafter called Compound 15)
[0071] (16)
2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-carbon-
itrile ethylene ketal (hereinafter called Compound 16) [0072] (17)
8-hydroxy-9-methoxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthry-
lene-10b-carbonitrile ethylene ketal (hereinafter called Compound
21) [0073] (18)
9-hydroxy-8-methoxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthry-
lene-10b-carbonitrile ethylene ketal (hereinafter called Compound
22)
[0074] The prophylactic and/or therapeutic agent for the
neurological disorder is desirably a compound that inhibits the
neuronal cell death as well as extends the neurites of the
remaining neuronal cells to normalize the signal transduction
between the nerve cells. The compound of general formula (1) has
the neuronal cell death inhibition effect and the neurite outgrowth
effect and therefore is useful as an active component in a medical
drug for preventing and/or treating the neurological disorders
caused by the neuronal cell death or the neurite atrophy.
Neurological disorders to be applied with the medical drug of the
present invention are not specifically limited. Examples thereof
include Alzheimer's disease, cerebrovascular dementia, senile
dementia, frontotemporal dementia, Lewy body dementia, Parkinson's
disease, Huntington's disease, neurogenic bladder, overactive
bladder, nervous pollakiuria, urge incontinence, reflex
incontinence, overflow incontinence, amyotrophic lateral sclerosis,
cerebral hemorrhage, cerebral infarction, brain tumor, brain
damage, spinal cord injury, Down syndrome, and hyperactivity
disorder.
[0075] As the active component in the medical drug provided by the
present invention, a substance selected from a group consisting of
a free compound of general formula (1), a physiologically
acceptable salt thereof, a hydrate thereof, and a solvate thereof
may be used. Generally, the medical drug of the present invention
is provided in a form of a pharmaceutical composition containing
the active component and formulation additives (such as a carrier
and a diluent).
[0076] The administration route of the medical drug of the present
invention is not specifically limited and may be any of oral
administration or parenteral administration (such as intramuscular
administration, intravenous administration, subcutaneous
administration, intraperitoneal administration, mucosal
administration through nasal cavity and the like, and inhalation
administration). The form of the medical drug of the present
invention is not specifically limited. Examples of the preparation
for oral administration include tablets, capsules, fine granules,
powders, granules, liquids, and syrups, and examples of the
preparation for parenteral administration include injections,
drops, suppositories, inhalations, transmucosal absorption agents,
transdermal absorption agents, nasal drops, ear drops, and eye
drops.
[0077] Each of the form, the formulation additives to be used, and
the formulation method for the medical drug of the present
invention can be properly selected by a person skilled in the art.
The dose of the medical drug of the present invention can be
properly selected by comprehensively considering the sex, the age
or weight, the severity of a symptom, the administration object
such as prevention or treatment, the presence or absence of
complicated symptoms, and the like of a patient. As for the dose,
typically, in the case of the oral administration for adults, about
0.05 mg to 2 g, preferably about 0.1 mg to 1 g may be administered
per day divided into 1 to 5 doses. In the case of the parenteral
administration for adults, about 0.01 mg to 1 g, preferably about
0.05 mg to 0.5 mg may be administered per day divided into 1 to 5
doses.
[0078] The present invention will be further described in detail
with reference to Examples and Reference Examples, but the present
invention is not limited thereto.
EXAMPLES
Reference Example 1
Synthesis of
5-hydroxy-1-[3-(furan-3-yl)-3-oxo-prop-1-yl]-benzocyclobutene-1-carbonitr-
ile ethylene ketal (Compound 18)
##STR00006##
[0080] Under an argon atmosphere, to a THF solution (50 mL) of
5-triisopropylsilyloxy-1-[3-(furan-3-yl)-3-oxo-prop-1-yl]-benzocyclobuten-
e-1-carbonitrile ethylene ketal (hereinafter called Compound 17)
(542 mg, 1.16 mmol) (Heterocycles 2004, 62, 207-211),
tetra-n-butylammonium fluoride (1 M THF solution, 1.27 mL, 1.27
mmol) was added dropwise at 0.degree. C., and the whole was stirred
at room temperature for 0.5 hour. The reacted solution was diluted
with a saturated aqueous ammonium chloride solution and extracted
with chloroform. The organic phase was dried over magnesium sulfate
and filtered, and then the solvent was removed by evaporation. The
residue was purified by silica gel column chromatography
(hexane:acetone=3:2) to produce
5-hydroxy-1-[3-(furan-3-yl)-3-oxo-prop-1-yl]-benzocyclobutene-1-carbonitr-
ile ethylene ketal (hereinafter called Compound 18) (362 mg,
quant.) as yellow oil.
[0081] .sup.1H-NMR (270 MHz, CDCl.sub.3) .delta.: 1.94-2.10 (2H,
m), 2.14-2.24 (2H, m), 3.14 (1H, d, J=14 Hz), 3.59 (1H, d, J=14
Hz), 3.85-4.05 (4H, m), 6.31 (1H, dd, J=1.0, 1.7 Hz), 6.60 (1H, m),
6.67 (1H, dd, J=2.3, 8.2 Hz), 6.99 (1H, d, J=8.2 Hz), 7.34-7.38
(2H, m); .sup.13C-NMR (67.5 MHz, CDCl.sub.3) .delta.: 31.5, 36.0,
41.4, 42.1, 63.6, 64.8, 106.9, 108.5, 111.1, 115.7, 121.8, 123.0,
127.1, 134.8, 140.1, 142.1, 143.6, 157.3; IR (neat): 3389, 2234
cm.sup.-1; MS (EI) m/z 311 (M.sup.+); HRMS (EI) Calcd for
C.sub.18H.sub.17NO.sub.4: 311.1158, Found: 311.1135.
Reference Example 2
Synthesis of
5-trifluoromethanesulfonyloxy-1-[3-(furan-3-yl)-3-oxo-prop-1-yl]-benzocyc-
lobutene-1-carbonitrile ethylene ketal (Compound 19)
##STR00007##
[0083] Under an argon atmosphere, to a dichloromethane solution (15
mL) of Compound 18 (362 mg, 1.16 mmol), pyridine (0.281 mL, 3.48
mmol) was added and stirred. To the mixture, a dichloromethane
solution (2 mL) of trifluoromethanesulfonic anhydride (0.390 mL,
2.32 mmol) was added dropwise at 0.degree. C., and the whole was
stirred for 0.5 hour. Water was added to stop the reaction, the
reacted solution was extracted with dichloromethane, the organic
phase was dried over magnesium sulfate and filtered, and then the
solvent was removed by evaporation. The residue was purified by
silica gel column chromatography (ethyl acetate:hexane=1:4) to
produce
5-trifluoromethanesulfonyloxy-1-[3-(furan-3-yl)-3-oxo-prop-1-yl]-benzocyc-
lobutene-1-carbonitrile ethylene ketal (hereinafter called Compound
19) (484 mg, 94%) as colorless oil.
[0084] .sup.1H-NMR (270 MHz, CDCl.sub.3) .delta.: 2.02-2.12 (2H,
m), 2.13-2.25 (2H, m), 3.28 (1H, d, J=15 Hz), 3.71 (1H, d, J=15
Hz), 3.78-4.00 (4H, m), 6.31-6.32 (1H, m), 7.09-7.10 (1H, m), 7.17
(1H, dd, J=2.3, 8.2 Hz), 7.28 (1H, d, J=8.2 Hz), 7.35-7.38 (2H, m);
.sup.13C-NMR (67.5 MHz, CDCl.sub.3) .delta.: 31.2, 36.1, 42.1,
42.2, 106.6, 108.4, 118.0, 118.7 (q, J=321 Hz), 120.4, 122.0,
124.0, 127.1, 140.1, 143.0, 143.6, 143.6, 150.4; IR (neat): 2236
cm.sup.-1; MS (EI) m/z 443 (M.sup.+); HRMS (EI) Calcd for
C.sub.19H.sub.16F.sub.3NO.sub.6S: 443.0650, Found: 443.0621.
Reference Example 3
Synthesis of
8-trifluoromethanesulfonyloxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-ace-
phenanthrylene-10b-carbonitrile ethylene ketal (Compound 6)
##STR00008##
[0086] Under an argon atmosphere, to a dichloromethane solution (3
mL) of Compound 5 (56.2 mg, 0.181 mmol) (J. Org. Chem. 2004, 69,
7989-7993), pyridine (0.044 mL, 0.542 mmol) was added and stirred.
To the mixture, a dichloromethane solution (1 mL) of
trifluoromethanesulfonic anhydride (0.061 mL, 0.361 mmol) was added
dropwise at 0.degree. C., and the whole was stirred at 0.degree. C.
for 0.5 hour. Water was added to stop the reaction, the reacted
solution was extracted with dichloromethane, the organic phase was
dried over magnesium sulfate and filtered, and then the solvent was
removed by evaporation. The residue was purified by silica gel
column chromatography (dichloromethane:hexane=10:1) to produce
Compound 6 (72.1 mg, 90%) as a white solid (mp 127 to 129.degree.
C.).
[0087] .sup.1H-NMR (270 MHz, CDCl.sub.3) .delta.: 1.69-1.87 (2H,
m), 2.53 (1H, td, J=4.6, 15 Hz), 2.78-2.86 (1H, m), 3.14 (1H, dd,
J=2.3, 16 Hz), 3.25 (1H, dd, J=3.3, 16 Hz), 3.78-3.99 (5H, m), 5.26
(1H, ddd, J=2.6, 3.3, 11 Hz), 5.94 (1H, d, J=2.0 Hz), 7.16-7.19
(2H, m), 7.33-7.37 (1H, m); .sup.13C-NMR (67.5 MHz, CDCl.sub.3)
.delta.: 29.6, 31.3, 33.9, 35.5, 51.1, 63.8, 6.4, 79.1, 104.3,
110.2, 118.7 (q, J=321 Hz), 120.1, 121.2, 123.2, 127.7, 131.6,
138.7, 143.2, 149.3; IR (KBr): 2231 cm.sup.-1; MS (EI): m/z 443
(M.sup.+); HRMS (EI) Calcd for C.sub.19H.sub.16F.sub.3NO.sub.6S:
443.0650, Found: 443.0664.
Reference Example 4
Synthesis of
8-(2,4,6-trifluorophenyl)-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephen-
anthrylene-10b-carbonitrile ethylene ketal (Compound 10)
##STR00009##
[0089] Under an argon atmosphere, into a 1,4-dioxane solution (3
mL) of Compound 6 (158 mg, 0.356 mmol), 2,4,6-trifluorophenylboric
acid (68.8 mg, 0.391 mmol), potassium bromide (46.5 mg, 0.391
mmol), tripotassium phosphate hexahydrate (142 mg, 0.533 mmol), and
tetrakis(triphenylphosphine)palladium (41.1 mg, 0.0356 mmol) were
mixed, and the mixture was heated and refluxed for 5 hours. After
being cooled, the reacted solution was diluted with dichloromethane
and washed with a saturated aqueous sodium hydrogen carbonate, then
the organic phase was dried over magnesium sulfate and filtered,
and then the solvent was removed by evaporation. The residue was
purified by silica gel column chromatography (ethyl
acetate:hexane=1:3) to produce Compound 10 (60.5 mg, 40%) as a
white solid (mp 136 to 137.degree. C.).
[0090] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.82-1.97 (2H,
m), 2.56 (1H, ddd, J=5.2, 13, 15 Hz), 2.89-2.96 (1H, m), 3.17 (1H,
dd, J=2.5, 16 Hz), 3.30 (1H, dd, J=3.6, 16 Hz), 3.84-4.07 (5H, m),
5.33 (1H, m), 6.01 (1H, d, J=1.4 Hz), 7.26-7.40 (5H, m);
.sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 29.9, 31.6, 34.1, 35.9,
51.4, 63.9, 65.5, 79.9, 104.8, 110.5, 122.2, 125.7, 127.5, 128.7,
130.6, 131.0, 135.5, 143.2; IR (KBr): 2228 cm.sup.-1; MS (EI) m/z
425 (M.sup.+); HRMS (EI) Calcd for C.sub.24H.sub.18F.sub.3NO.sub.3:
425.1239, Found: 425.1262.
Reference Example 5
Synthesis of
8-phenyl-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-ca-
rbonitrile ethylene ketal (Compound 9)
##STR00010##
[0092] Under an argon atmosphere, into a 1,4-dioxane solution (3
mL) of.
[0093] Compound 19 (203 mg, 0.457 mmol), phenylboric acid (61.3 mg,
0.503 mmol), potassium bromide (59.9 mg, 0.503 mmol), tripotassium
phosphate hexahydrate (183 mg, 0.686 mmol), and
tetrakis(triphenylphosphine)palladium (52.9 mg, 0.0457 mmol) were
mixed, and the mixture was heated and refluxed for 1 hour. After
being cooled, the reacted solution was diluted with dichloromethane
and washed with a saturated aqueous sodium hydrogen carbonate, then
the organic phase was dried over magnesium sulfate and filtered,
and then the solvent was removed by evaporation. The residue was
subjected to silica gel column chromatography (ethyl
acetate:hexane=1:4) to produce a coupled compound (161 mg) as a
white solid. The compound was successively dissolved in
o-dichlorobenzene (25 mL), the solution was heated and refluxed for
1 hour, and then the solvent was removed by evaporation. The
residue was purified by silica gel column chromatography (ethyl
acetate:hexane=1:3) to produce Compound 9 (104 mg, a two-step yield
of 61%) as a white solid (mp 179.degree. C.).
[0094] .sup.1H-NMR (270 MHz, CDCl.sub.3) .delta.: 1.83-2.00 (2H,
m), 2.57 (1H, ddd, J=4.9, 13, 15 Hz), 2.90-2.97 (1H, m), 3.22 (1H,
dd, J=2.3, 16 Hz), 3.34 (1H, dd, J=3.0, 16 Hz), 3.80-4.01 (5H, m),
5.34 (1H, ddd, J=2.3, 3.3, 11 Hz), 6.03 (1H, d, J=1.7 Hz),
7.29-7.59 (8H, m); .sup.13C-NMR (67.5 MHz, CDCl.sub.3) .delta.:
29.7, 31.4, 34.0, 35.5, 51.1, 63.7, 65.3, 79.7, 104.6, 110.3,
122.0, 126.1, 127.1, 127.6, 128.8, 129.1, 129.9, 135.8, 140.2,
141.5, 1432; IR (KBr): 2226 cm.sup.-1; MS (EI) m/z 371 (M.sup.+);
HRMS (EI) Calcd for C.sub.24H.sub.21NO.sub.3: 371.1522, Found:
371.1518.
Reference Example 6
Synthesis of
7-[3-(tetrahydropyran-2-yloxy)-propyl]-4,6-dimethoxy-3-oxobicyclo[4.2.0]o-
cta-1,4-diene-7-carbonitrile (Compound 24)
##STR00011##
[0096] Under an argon atmosphere, to a MeOH solution (20 mL) of
Compound 23 (541 mg, 1.88 mmol) (Heterocycles 2004, 62, 207-211),
PhI(OAc).sub.2 (1.33 g, 4.14 mmol) was added, the whole was stirred
at room temperature for 30 minutes, and then the reaction was
stopped by a saturated aqueous NaHCO.sub.3 solution. The organic
phase extracted with CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and
filtered, and then the solvent was removed by evaporation. The
residue was purified by silica gel column chromatography
(AcOEt:hexane=1:1) to produce Compound 24 (333 mg, 959 .mu.mol,
51%) as yellow oil.
[0097] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.52-2.17 (10H,
m), 3.20 (1H, dd, J=1.6 Hz J=17.9 Hz), 3.37 (6H, d, J=1.37 Hz),
3.43-3.54 (2H, m), 3.60 (1H, dd, J=1.9 Hz J=17.9 Hz), 3.79-3.87
(2H, m), 4.57 (1H, s), 5.92 (1H, dd, J=1.9 Hz J=3.3 Hz), 6.33 (1H,
s); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 19.71, 19.76, 25.37,
26.03, 30.66, 34, 17, 41.34, 41.92, 50.19, 50.35, 62.63, 66.13,
93.55, 99.00, 119.49, 126.80, 142.75, 155.18, 192.99; IR (neat):
1682, 2236 cm.sup.-1; MS (EI): m/z 347 (M.sup.+); HRMS (EI): calcd
for C.sub.19H.sub.25NO.sub.5 347.1733, found 347.1684.
Reference Example 7
Synthesis of
3-hydroxy-4-methoxy-7-[3-(tetrahydropyran-2-yloxy)-propyl]bicyclo[4.2.0]o-
cta-1,3,5-triene-7-carbonitrile (Compound 25)
##STR00012##
[0099] To a mixed solution of Compound 24 (464 mg, 1.34 mmol), AcOH
(700 .mu.L), THF (7.0 mL), and H.sub.2O (7.0 mL), Zn (438 mg, 6.70
mmol) was added, the whole was stirred at room temperature for 30
minutes, and then the reaction was stopped by a saturated aqueous
NaHCO.sub.3 solution. The organic phase extracted with
CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and filtered, and then
the solvent was removed by evaporation. The residue was purified by
silica gel column chromatography (AcOEt:hexane=1:1) to produce
Compound 25 (365 mg, 1.15 mmol, 85%) as yellow oil.
[0100] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.54-2.03 (10H,
m), 3.19 (1H, d, J=13.7 Hz), 3.44-3.52 (2H, m), 3.62 (1H, d, J=13.7
Hz), 3.77-3.85 (2H, m), 3.89 (3H, s), 4.57 (1H, t, J=3.57 Hz), 5.83
(1H, s), 6.74 (1H, s), 6.76 (1H, d, J=1.9 Hz); .sup.13C-NMR (75
MHz, CDCl.sub.3) .delta.: 19.74, 25.48, 26.71, 30.74, 34.55, 42.05,
42.25, 56.39, 62.52, 66.65, 98.91, 104.83, 110.35, 121.98, 133.31,
134.10, 147.27, 147.79; IR (neat): 2230, 3418 cm.sup.-1; MS (EI):
m/z 317 (M.sup.+); HRMS (EI): calcd for C.sub.18H.sub.23NO.sub.4
317.1627, found 317.1604.
Reference Example 8
Synthesis of
7-[3-(tetrahydropyran-2-yloxy)-propyl]-3-triisopropylsilanyloxy-4-methoxy-
-bicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound 26)
##STR00013##
[0102] Under an argon atmosphere, to a CH.sub.2Cl.sub.2 solution
(8.0 mL) of Compound 25 (320 mg, 1.01 mmol), Et.sub.3N (238 .mu.L,
1.72 mmol) and TIPS-OTf (409 .mu.L, 1.52 mmol) were added at
0.degree. C., and the whole was stirred at room temperature for 1
hour. Then, the reacted solution was diluted with CH.sub.2Cl.sub.2
and successively washed with a 3% aqueous HCl solution and a
saturated aqueous NaHCO.sub.3 solution. Then the organic phase was
dried over MgSO.sub.4 and filtered, and then the solvent was
removed by evaporation. The residue was purified by silica gel
column chromatography (AcOEt:hexane=1:6) to produce Compound 26
(378 mg, 798 .mu.mol, 79%) as colorless oil.
[0103] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=7.1 Hz), 1.17-1.29 (3H, m), 1.56-2.04 (10H, m), 3.16 (1H, d,
J=13.5 Hz), 3.43-3.50 (2H, m), 3.60 (1H, d, J=13.5 Hz), 3.78 (3H,
s), 3.79-3.84 (2H, m), 4.57 (1H, s), 6.67 (1H, s), 6.72 (1H, s);
.sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.88, 17.91, 19.63,
25.42, 26.57, 30.66, 34.46, 41.87, 42.02, 55.69, 62.36, 66.63,
98.79, 105.91, 115.91, 122.00, 132.16, 135.26, 147.56, 151.96; IR
(neat): 2231 cm.sup.-1; MS (EI): m/z 473 (M.sup.+); HRMS (EI):
calcd for C.sub.27H.sub.43NO.sub.4Si 473.2961, found 473.2987.
Reference Example 9
Synthesis of
7-(3-hydroxypropyl)-3-triisopropylsilanyloxy-4-methoxy-bicyclo[4.2.0]octa-
-1,3,5-triene-7-carbonitrile (Compound 27)
##STR00014##
[0105] Under an argon atmosphere, to an EtOH solution (7.0 mL) of
Compound 26 (329 mg, 695 .mu.mol), PPTS (35.0 mg, 139 .mu.mol) was
added, the whole was heated and stirred at 75.degree. C. for 2
hours, and then the reaction was stopped by a saturated aqueous
NaHCO.sub.3 solution. The organic phase extracted with
CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and filtered, and then
the solvent was removed by evaporation. The residue was purified by
silica gel column chromatography (AcOEt:hexane=1:1) to produce
Compound 27 (266 mg, 683 .mu.mol, 98%) as colorless oil.
[0106] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.17-1.29 (3H, m), 1.84-1.92 (2H, m), 1.97-2.06 (2H, m),
2.17 (1H, s), 3.16 (1H, d, J=13.5 Hz), 3.61 (1H, d, J=13.5 Hz),
3.73 (2H, t, J=6.0 Hz), 3.77 (3H, s), 6.67 (1H, s), 6.71 (1H, s);
.sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.86, 17.91, 29.32,
34.01, 41.79, 42.04, 55.72, 61.90, 105.86, 115.93, 122.00, 132.14,
135.15, 147.61, 151.98; IR (neat): 2232, 3446 cm.sup.-1; MS (EI):
m/z 389 (M.sup.+); HRMS (EI): calcd for C.sub.22H.sub.35NO.sub.3Si
389.2386, found 389.2405.
Reference Example 10
Synthesis of
3-triisopropylsilanyloxy-4-methoxy-7-(3-oxopropyl)bicyclo[4.2.0]octa-1,3,-
5-triene-7-carbonitrile (Compound 28)
##STR00015##
[0108] Under an argon atmosphere, to a mixed solution of Compound
27 (97.4 mg, 250 .mu.mol), DMSO (2.0 mL), and Et.sub.3N (2.0 mL),
Py-SO.sub.3 (199 mg, 1.25 mmol) was added at room temperature, the
whole was stirred, and then the reaction was stopped by H.sub.2O.
The organic phase extracted with CH.sub.2Cl.sub.2 was successively
washed with H.sub.2O and a saturated aqueous NaCl solution, dried
over MgSO.sub.4, and filtered, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:2) to produce Compound 28 (89.7 mg,
231 .mu.mol, 92%) as yellow oil.
[0109] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.17-1.29 (3H, m), 2.23-2.32 (2H, m), 2.74-2.81 (2H, m),
3.16 (1H, d, J=13.7 Hz), 3.62 (1H, d, J=13.7 Hz), 3.78 (3H, s),
6.68 (1H, s), 6.69 (1H, s), 9.82 (1H, s); .sup.13C-NMR (75 MHz,
CDCl.sub.3) .delta.: 12.90, 17.95, 29.63, 40.42, 41.05, 41.83,
55.77, 105.78, 116.06, 121.41, 131.98, 134.23, 147.98, 152.24,
199.58; IR (neat): 1730, 2231 cm.sup.-1; MS (EI): m/z 387
(M.sup.+); HRMS (EI): calcd for C.sub.22H.sub.33NO.sub.3Si
387.2230, found 387.2274.
Reference Example 11
Synthesis of
7-[3-(furan-3-yl)-3-hydroxypropyl]-3-triisopropylsilanyloxy-4-methoxy-bic-
yclo[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound 29)
##STR00016##
[0111] Under an argon atmosphere, to an Et.sub.2O solution (2.0 mL)
of 3-bromofuran (37.4 mL, 416 .mu.mol), n-BuLi (1.6 M THF solution,
220 .mu.l, 352 .mu.mol) was added dropwise at -78.degree. C., and
the whole was stirred for 30 minutes. Then an Et.sub.2O solution
(2.0 mL) of Compound 28 (124 mg, 320 .mu.mol) was added dropwise to
the reacted solution at -78.degree. C., the temperature was raised
to 0.degree. C., the whole was stirred for 1.5 hours, and then the
reaction was stopped by a saturated aqueous NH.sub.4Cl solution.
The organic phase extracted with Et.sub.2O was dried over
MgSO.sub.4 and filtered, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:3) to produce Compound 29 (47.7 mg,
105 .mu.mol, 33%) as yellow oil.
[0112] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.12-1.29 (3H, m), 1.84 (1H, s), 1.88-2.14 (4H, m), 3.14
(1H, d, J=13.5 Hz), 3.59 (1H, d, J=13.5 Hz), 3.77 (3H, s), 6.41
(1H, dt, J=1.1 Hz J=2.5 Hz), 6.69 (2H, d, J=10.7 Hz), 7.40 (2H, t,
J=1.6 Hz); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.92, 17.98,
33.59, 33.66, 34.19, 41.71, 41.99, 42.05, 55.82, 66.37, 105.93,
108.15, 116.03, 121.98, 128.37, 132.16, 135.07, 139.05, 143.51,
147.72, 152.09; IR (neat): 2231, 3445 cm.sup.-1; MS (EI): m/z 455
(M.sup.+); HRMS (EI): calcd for C.sub.26H.sub.37NO.sub.3Si
455.2492, found 455.2541.
Reference Example 12
Synthesis of
7-[3-(furan-3-yl)-3-oxopropyl]-3-triisopropylsilanyloxy-4-methoxy-bicyclo-
[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound 30)
##STR00017##
[0114] Under an argon atmosphere, to a mixed solution of Compound
29 (41.3 mg, 90.6 .mu.mol), DMSO (1.0 mL), and Et.sub.3N (1.0 mL),
Py-SO.sub.3 (83.6 mg, 525 .mu.mol) was added at room temperature,
the whole was stirred, and then the reaction was stopped by
H.sub.2O. The organic phase extracted with CH.sub.2Cl.sub.2 was
successively washed with H.sub.2O and a saturated aqueous NaCl
solution, dried over MgSO.sub.4, and filtered, and then the solvent
was removed by evaporation. The residue was purified by silica gel
column chromatography (AcOEt:hexane=1:4) to produce Compound 30
(31.4 mg, 69.2 .mu.mol, 76%) as brown oil.
[0115] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.21-1.29 (3H, m), 2.37 (2H, dd, J=1.4 Hz J=7.4 Hz),
2.95-3.13 (2H, m), 3.19 (1H, d, J=13.7 Hz), 3.62 (1H, d, J=13.7
Hz), 3.77 (3H, s), 6.69 (2H, d, J=6.0 Hz), 6.77 (1H, dd, J=0.8 Hz
J=1.8 Hz), 7.45 (1H, t, J=1.8 Hz), 8.07 (1H, dd, J=0.8 Hz J=1.4
Hz); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.93, 17.96,
31.49, 36.72, 41.31, 41.86, 55.81, 105.86, 108.42, 116.07, 127.22,
132.08, 134.50, 144.26, 147.14, 147.93, 152.22, 192.49; IR (neat):
1683, 2230 cm.sup.-1; MS (EI): m/z 454 (M.sup.+); HRMS (EI): calcd
for C.sub.26H.sub.36NO.sub.4Si 454.2414, found 454.2395.
Reference Example 13
Synthesis of
7-[2-(2-furan-3-yl-[1,3]dioxolane-2-yl)ethyl]-3-triisopropylsilanyloxy-4--
methoxy-bicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound
31)
##STR00018##
[0117] Under an argon atmosphere, to a benzene solution (5.0 mL) of
Compound 30 (83.7 mg, 185 .mu.mol), ethylene glycol (103 .mu.L,
1.85 mmol) and TsOH (3.5 mg, 18.5 .mu.mol) were added at room
temperature, the whole was heated and refluxed for 20 hours using a
Dean-Stark apparatus, and then the reaction was stopped by a
saturated aqueous NaHCO.sub.3 solution. The organic phase extracted
with CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and filtered, and
then the solvent was removed by evaporation. The residue was
purified by silica gel column chromatography (AcOEt:hexane=1:3) to
produce Compound 31 (74.8 mg, 150 .mu.mol, 81%) as yellow oil.
[0118] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.12-1.31 (3H, m), 1.99-2.27 (4H, m), 3.12 (1H, d,
J=13.5 Hz), 3.57 (1H, d, J=13.5 Hz), 3.77 (3H, s), 3.86-4.03 (4H,
m), 6.33 (1H, q, J=0.8 Hz), 6.66 (1H, s), 6.69 (1H, s), 7.36 (1H,
t, J=1.6 Hz), 7.39 (1H, t, J=0.8 Hz); .sup.13C-NMR (75 MHz,
CDCl.sub.3) .delta.: 12.93, 17.98, 31.62, 36.00, 41.53, 41.89,
55.81, 64.80, 64.83, 105.88, 106.77, 108.42, 115.98, 127.11,
132.16, 135.09, 139.92, 143.37, 147.67, 152.06; IR (neat): 2231
cm.sup.-1; MS (EI): m/z 497 (M.sup.+); HRMS (EI): calcd for
C.sub.28H.sub.39NO.sub.5Si 497.25298, found 497.2569.
Reference Example 14
Synthesis of
8-triisopropylsilanyloxy-9-methoxy-3-[1,3]dioxolane-2,3,5a,10c-tetrahydro-
-1H,6H-5-oxa-acephenanthrylene-10b-carbonitrile (Compound 32)
##STR00019##
[0120] Under an argon atmosphere, an o-dichlorobenzene solution
(2.0 mL) of Compound 31 (27.6 mg, 55.5 .mu.mol) was heated and
refluxed for 1.5 hours, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:3) to produce Compound 32 (17.7 mg,
35.6 .mu.mol, 64%) as colorless oil.
[0121] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.13-1.30 (3H, m), 1.83-1.98 (2H, m), 2.53 (1H, m), 2.84
(1H, td, J=3.6 Hz J=14.6 Hz), 3.00 (1H, dd, J=2.2 Hz J=15.9 Hz),
3.18 (1H, dd, J=3.6 Hz J=15.9 Hz), 3.79 (3H, s), 3.84-3.95 (4H, m),
3.96-4.05 (1H, m), 5.27 (1H, ddd, J=2.2 Hz J=3.6 Hz J=10.4 Hz),
6.00 (1H, d, J=2.2 Hz), 6.76 (1H, s), 6.79 (1H, s); .sup.13C-NMR
(75 MHz, CDCl.sub.3) .delta.: 12.98, 17.99, 30.00, 31.62, 33.17,
35.53, 51.18, 55.92, 63.70, 65.32, 79.93, 104.60, 110.22, 122.24,
122.40, 122.87, 127.79, 143.01, 145.71, 149.87; IR (neat): 2228
cm.sup.-1; MS (EI): m/z 497 (M.sup.+); HRMS (EI): calcd for
C.sub.28H.sub.53NO.sub.5Si 497.3662, found 497.3624.
Reference Example 15
Synthesis of
4-hydroxy-3-methoxy-7-[3-(tetrahydropyran-2-yloxy)-propyl]bicyclo[4.2.0]o-
cta-1,3,5-triene-7-carbonitrile (Compound 34)
##STR00020##
[0123] Under an argon atmosphere, to a MeOH solution (60 mL) of
Compound 33 (3.03 g, 10.5 mmol) (Heterocycles 2004, 62, 207-211),
PhI(OAc).sub.2 (7.44 g, 23.1 mmol) was added, the whole was stirred
at room temperature for 30 minutes, and then the reaction was
stopped by a saturated aqueous NaHCO.sub.3 solution. The organic
phase extracted with CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and
filtered, and then the solvent was removed by evaporation. The
residue was separated by silica gel column chromatography
(AcOEt:hexane=1:1), and the solvent was removed by evaporation. To
a mixed solution of the residue, AcOH (6.0 mL), THF (60.0 mL), and
H.sub.2O (60.0 mL), Zn (1.22 mg, 18.7 mmol) was added, the whole
was stirred at room temperature for 30 minutes, and then the
reaction was stopped by a saturated aqueous NaHCO.sub.3 solution.
The organic phase extracted with CH.sub.2Cl.sub.2 was dried over
MgSO.sub.4 and filtered, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:2) to produce Compound 34 (447 mg,
1.41 mmol, 13%) as yellow oil.
[0124] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.48-2.09 (10H,
m), 3.18 (1H, d, J=13.5 Hz), 3.41-3.51 (2H, m), 3.61 (1H, d, J=13.5
Hz), 3.74-3.92 (2H, m), 3.87 (3H, s), 4.57 (1H, br), 5.83 (1H, s),
6.68 (1H, s), 6.80 (1H, d, J=1.4 Hz); .sup.13C-NMR (75 MHz,
CDCl.sub.3) .delta.: 19.63, 25.45, 26.59, 30.68, 34.45, 41.99,
42.25, 56.29, 62.36, 66.63, 98.81, 106.75, 108.36, 121.80, 131.70,
135.36, 1465.1, 148.48; IR (neat): 3449 cm.sup.-1; MS (EI): m/z 317
(M.sup.+).
Reference Example 16
Synthesis of
7-[3-(tetrahydropyran-2-yloxy)-propyl]-4-triisopropylsilanyloxy-3-methoxy-
-bicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound 35)
##STR00021##
[0126] Under an argon atmosphere, to a CH.sub.2Cl.sub.2 solution
(12.0 mL) of Compound 34 (493 mg, 1.55 mmol), Et.sub.3N (366 .mu.L,
2.64 mmol) and TIPS-OTf (626 .mu.L, 2.33 mmol) were added at
0.degree. C., and the whole was stirred at room temperature for 45
minutes. Then, the reacted solution was diluted with
CH.sub.2Cl.sub.2 and subsequently washed with a 3% aqueous HCl
solution and a saturated aqueous NaHCO.sub.3 solution, the organic
phase was dried over MgSO.sub.4 and filtered, and then the solvent
was removed by evaporation. The residue was purified by silica gel
column chromatography (AcOEt:hexane=1:4) to produce Compound 35
(666 mg, 1.41 mmol, 91%) as yellow oil.
[0127] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.15-1.29 (3H, m), 1.53-2.04 (10H, m), 3.18 (1H, d,
J=13.7 Hz), 3.41-3.52 (2H, m), 3.61 (1H, d, J=13.7 Hz), 3.79-3.85
(2H, m), 3.77 (3H, s), 4.57 (1H, br), 6.65 (1H, s), 6.74 (1H, d,
J=1.6 Hz); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.96, 17.99,
19.69, 25.50, 26.62, 30.73, 34.56, 41.83, 42.21, 55.69, 62.39,
66.65, 98.79, 107.92, 113.95, 122.03, 133.11, 134.60, 146.38,
153.22; IR (neat): 2231 cm.sup.-1; MS (EI): m/z 473 (M.sup.+).
Reference Example 17
Synthesis of
7-(3-hydroxypropyl)-4-triisopropylsilanyloxy-3-methoxy-bicyclo[4.2.0]octa-
-1,3,5-triene-7-carbonitrile (Compound 36)
##STR00022##
[0129] Under an argon atmosphere, to an EtOH solution (10.0 mL) of
Compound 35 (666 mg, 1.41 mmol), PPTS (70.9 mg, 282 .mu.mol) was
added, the whole was heated and stirred at 75.degree. C. for 2
hours, and then the reaction was stopped by a saturated aqueous
NaHCO.sub.3 solution. The organic phase extracted with
CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and filtered, and then
the solvent was removed by evaporation. The residue was purified by
silica gel column chromatography (AcOEt:hexane=1:2) to produce
Compound 36 (576 mg, 1.48 .mu.mol, 105%) as colorless oil.
[0130] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.6 Hz), 1.12-1.29 (3H, m), 1.45 (1H, br), 1.80-1.90 (2H, m),
1.96-2.04 (2H, m), 3.17 (1H, d, J=13.5 Hz), 3.62 (1H, d, J=13.5
Hz), 3.72 (2H, t, J=6.0 Hz), 3.77 (3H, s), 6.65 (1H, s), 6.73 (1H,
s); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.96, 17.99, 29.42,
34.08, 41.75, 42.26, 55.71, 62.13, 107.95, 113.86, 121.98, 133.06,
134.50, 146.43, 153.27; IR (neat): 2232, 3437 cm.sup.-1; MS (EI):
m/z 389 (M.sup.+).
Reference Example 18
Synthesis of
4-triisopropylsilanyloxy-3-methoxy-7-(3-oxopropyl)bicyclo[4.2.0]octa-1,3,-
5-triene-7-carbonitrile (Compound 37)
##STR00023##
[0132] Under an argon atmosphere, to a mixed solution of Compound
36 (123 mg, 316 .mu.mol), DMSO (2.0 mL), and Et.sub.3N (2.0 mL),
Py-SO.sub.3 (251 mg, 1.58 mmol) was added at room temperature, the
whole was stirred, and then the reaction was stopped by H.sub.2O.
The organic phase extracted with CH.sub.2Cl.sub.2 was successively
washed with H.sub.2O and a saturated aqueous NaCl solution, dried
over MgSO.sub.4, and filtered, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:3) to produce Compound 37 (94.9 mg,
245 .mu.mol, 78%) as yellow oil.
[0133] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.07 (18H, d,
J=6.9 Hz), 1.17-1.31 (3H, m), 2.16-2.35 (2H, m), 2.75 (2H, t, J=7.7
Hz), 3.17 (1H, d, J=13.5 Hz), 3.63 (1H, d, J=13.5 Hz), 3.76 (3H,
s), 6.66 (1H, s), 6.71 (1H, s), 9.81 (1H, s); .sup.13C-NMR (75 MHz,
CDCl.sub.3) .delta.: 12.90, 17.93, 29.56, 40.39, 40.95, 41.99,
55.61, 107.90, 113.71, 121.33, 132.82, 133.47, 146.54, 153.50,
199.48; IR (neat): 1726, 2231 cm.sup.-1; MS (EI): m/z 387
(M.sup.+).
Reference Example 19
Synthesis of
7-[3-(furan-3-yl)-3-hydroxypropyl]-4-triisopropylsilanyloxy-3-methoxy-bic-
yclo[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound 38)
##STR00024##
[0135] Under an argon atmosphere, to an Et.sub.2O solution (2.0 mL)
of 3-bromofuran (49.4 mL, 549 .mu.mol), n-BuLi (1.6 M THF solution,
298 .mu.L, 476 .mu.mol) was added dropwise at -78.degree. C., and
the whole was stirred for 30 minutes. Then, an Et.sub.2O solution
(2.0 mL) of Compound 37 (142 mg, 366 .mu.mol) was added dropwise to
the reacted solution at -78.degree. C., the temperature was raised
to 0.degree. C., the whole was stirred for 1.5 hours, and then the
reaction was stopped by a saturated aqueous NH.sub.4Cl solution.
The organic phase extracted with Et.sub.2O was dried over
MgSO.sub.4 and filtered, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:3) to produce Compound 38 (81.6 mg,
179 .mu.mol, 49%) as colorless oil.
[0136] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.9 Hz), 1.13-1.29 (3H, m), 1.84-2.17 (5H, m), 3.15 (1H, dd,
J=3.3 Hz J=13.7 Hz), 3.61 (1H, d, J=13.7 Hz), 3.77 (3H, s),
4.70-4.74 (1H, m), 6.38-6.40 (1H, m), 6.65 (1H, s), 6.73 (1H, s),
7.39 (2H, t, J=1.6 Hz); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.:
12.96, 17.99, 33.67, 34.27, 41.62, 42.17, 55.71, 66.44, 107.97,
108.11, 113.83, 121.91, 128.37, 133.05, 134.41, 139.00, 143.51,
146.43, 153.30; IR (neat): 2232, 3478 cm.sup.-1; MS (EI): m/z 455
(M)).
Reference Example 20
Synthesis of
7-[3-(furan-3-yl)-3-oxopropyl]-4-triisopropylsilanyloxy-3-methoxy-bicyclo-
[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound 39)
##STR00025##
[0138] Under an argon atmosphere, to a mixed solution of Compound
38 (167 mg, 367 .mu.mol), DMSO (2.0 mL), and Et.sub.3N (2.0 mL),
Py-SO.sub.3 (293 mg, 1.84 mmol) was added at room temperature, the
whole was stirred, and then the reaction was stopped by H.sub.2O.
The organic phase extracted with CH.sub.2Cl.sub.2 was successively
washed with H.sub.2O and a saturated aqueous NaCl solution, dried
over MgSO.sub.4, and filtered, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:4) to produce Compound 39 (126 mg,
278 .mu.mol, 76%) as yellow oil.
[0139] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.07 (18H, d,
J=7.1 Hz), 1.12-1.28 (3H, m), 2.32-2.38 (2H, m), 2.99-3.05 (2H, m),
3.20 (1H, d, J=13.7 Hz), 3.63 (1H, d, J=13.7 Hz), 3.76 (3H, s),
6.66 (1H, s), 6.72 (1H, s), 6.76 (1H, dd, J=0.8 Hz J=1.9 Hz), 7.44
(1H, dd, J=1.4 Hz J=1.9 Hz), 8.06 (1H, dd, J=0.8 Hz J=1.4 Hz);
.sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.90, 17.93, 31.46,
36.71, 41.21, 42.04, 55.63, 107.92, 108.39, 113.71, 121.59, 127.19,
132.92, 133.71, 144.21, 146.49, 147.07, 153.43, 192.41; Lit (neat):
1683, 2231 cm.sup.-1; MS (EI): m/z 453 (M.sup.+).
Reference Example 21
Synthesis of
7-[2-(2-furan-3-yl-[1,3]dioxolane-2-yl)ethyl]-4-triisopropylsilanyloxy-3--
methoxy-bicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile (Compound
40)
##STR00026##
[0141] Under an argon atmosphere, to a benzene solution (7.0 mL) of
Compound 39 (119 mg, 262 .mu.mol), ethylene glycol (147 .mu.L, 2.62
mmol) and TsOH (5.0 mg, 26.2 .mu.mol) were added at room
temperature, the whole was heated and refluxed for 13 hours using a
Dean-Stark apparatus, and then the reaction was stopped by a
saturated aqueous NaHCO.sub.3 solution. The organic phase extracted
with CH.sub.2Cl.sub.2 was dried over MgSO.sub.4 and filtered, and
then the solvent was removed by evaporation. The residue was
purified by silica gel column chromatography (AcOEt:hexane=1:3) to
produce Compound 40 (114 mg, 229 .mu.mol, 87%) as yellow oil.
[0142] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.6 Hz), 1.19-1.29 (3H, m), 1.97-2.19 (4H, m), 3.13 (1H, d,
J=13.5 Hz), 3.58 (1H, d, J=13.5 Hz), 3.76 (3H, s), 3.86-4.01 (4H,
m), 6.31 (1H, dd, J=1.1 Hz J=1.6 Hz), 6.63 (1H, s), 6.71 (1H, s),
7.36 (1H, t, J=1.6 Hz), 7.37 (1H, dd, J=1.1 Hz J=1.6 Hz);
.sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.93, 17.96, 31.60,
35.97, 41.37, 42.02, 55.66, 64.78, 106.74, 107.92, 108.37, 113.84,
121.80, 127.11, 133.01, 134.32, 139.87, 143.32, 146.35, 153.24; IR
(neat): 2231 cm.sup.-1; MS (EI): m/z 497 (M.sup.+).
Reference Example 22
Synthesis of
9-triisopropylsilanyloxy-8-methoxy-3-[1,3]dioxolane-2,3,5a,10c-tetrahydro-
-1H,6H-5-oxa-acephenanthrylene-10b-carbonitrile (Compound 41)
##STR00027##
[0144] Under an argon atmosphere, an o-dichloro-benzene solution
(2.0 mL) of Compound 40 (68.8 mg, 138 .mu.mol) was heated and
refluxed for 1 hour, and then the solvent was removed by
evaporation. The residue was purified by silica gel column
chromatography (AcOEt:hexane=1:3) to produce Compound 41 (44.6 mg,
89.6 .mu.mol, 65%) as a white solid (mp 131 to 133.degree. C.).
[0145] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.08 (18H, d,
J=6.6 Hz), 1.12-1.30 (3H, m), 1.79-1.95 (2H, m), 2.45-2.56 (1H, m),
2.75 (1H, dt, J=3.6 Hz J=14.6 Hz), 3.05 (1H, dd, J=2.2 Hz J=15.9
Hz), 3.22 (1H, dd, J=3.6 Hz J=15.9 Hz), 3.81 (3H, s), 3.84-3.94
(4H, m), 3.95-4.05 (1H, m), 5.28 (1H, ddd, J=2.2 Hz J=3.6 Hz J=10.7
Hz), 6.01 (1H, d, J=2.2 Hz), 6.75 (1H, s), 6.82 (1H, s);
.sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 12.96, 17.96, 29.92,
31.34, 33.53, 35.27, 51.03, 55.42, 63.68, 65.32, 79.93, 104.59,
110.43, 113.79, 117.85, 122.19, 122.22, 128.13, 142.82, 144.42,
150.67; IR (neat): 2358 cm.sup.-1; MS (EI): m/z 497 (M.sup.+).
Example 1
Synthesis of
9-hydroxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenanthrylene-10b-c-
arbonitrile ethylene ketal (Compound 11)
##STR00028##
[0147] Under an argon atmosphere, to a THF solution (2.0 mL) of
8-triisopropylsilyloxy-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3-oxo-acephenant-
hrylene-10b-carbonitrile ethylene ketal (hereinafter called
Compound 20) (19 mg, 0.041 mmol) (Bioorg. Med. Chem. 2007, 15,
424-432), tetra-n-butylammonium fluoride (1 M THF solution, 0.053
mL, 0.053 mmol) was added dropwise at 0.degree. C., and then the
whole was stirred at room temperature for 1.5 hours. The reacted
solution was diluted with a saturated aqueous ammonium chloride
solution and extracted with dichloromethane, the organic phase was
dried over magnesium sulfate and filtered, and then the solvent was
removed by evaporation. The residue was purified by silica gel
column chromatography (hexane:ethyl acetate=1:1) to produce
Compound 11 (12 mg, 95%) as a white solid (mp 61 to 63.degree.
C.).
[0148] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.82-1.98 (2H,
m), 2.47-2.58 (1H, m), 2.80 (1H, td, J=3.3, 15 Hz), 3.11 (1H, dd,
J=2.2, 16 Hz), 3.18 (1H, dd, J=3.3, 16 Hz), 3.87-3.96 (4H, m),
3.98-4.04 (1H, m), 5.29 (1H, td, J=2.8, 11 Hz), 6.03 (1H, d, J=1.9
Hz), 6.74 (1H, dd, J=2.5, 8.2 Hz), 6.86 (1H, d, J=2.2 Hz), 7.12
(1H, d, J=8.2 Hz); .sup.13C-NMR (75 MHz, CDCl.sub.3) .delta.: 29.7,
31.5, 32.9, 35.8, 51.0, 63.7, 65.3, 79.8, 104.6, 110.2, 113.1,
115.5, 121.8, 127.0, 131.3, 132.0, 143.0, 155.0; IR (KBr): 3396,
2232 cm.sup.-1; MS (EI): m/z 311 (M.sup.+); HRMS (EI): calcd for
C.sub.18H.sub.17NO.sub.4: 311.1158, Found: 311.1152.
Example 2
Synthesis of
8-hydroxy-9-methoxy-3-[1,3]dioxolane-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3--
oxo-acephenanthrylene-10b-carbonitrile ethylene ketal (Compound
21)
##STR00029##
[0150] Under an argon atmosphere, to a THF solution (2.0 mL) of
Compound 32 (17.7 mg, 35.6 .mu.mol), TBAF (1.0 M THF solution, 46.3
.mu.L, 46.3 .mu.mol) was added dropwise at 0.degree. C., the whole
was stirred at room temperature for 40 minutes, and then the
reaction was stopped by a saturated aqueous NH.sub.4Cl solution.
The organic phase extracted with CH.sub.2Cl.sub.2 was washed with
H.sub.2O, dried over MgSO.sub.4, and filtered, and then the solvent
was removed by evaporation. The residue was purified by silica gel
column chromatography (AcOEt:hexane=1:1) to produce Compound 21
(10.2 mg, 29.9 .mu.mol, 84%) as colorless oil.
[0151] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.84-1.97 (2H,
m), 2.55 (1H, ddd, J=5.5 Hz J=12.4 Hz J=14.8 Hz), 2.84 (1H, J=3.3
Hz J=14.8 Hz), 3.05 (1H, dd, J=2.2 Hz J=15.9 Hz), 3.19 (1H, dd,
J=3.6 Hz J=15.9 Hz), 3.88 (3H, s), 3.82-3.95 (4H, m), 3.97-4.06
(1H, m), 5.28 (1H, ddd, J=2.2 Hz J=3.6 Hz J=10.4 Hz), 5.66 (1H, s),
6.04 (1H, d, J=2.2 Hz), 6.77 (1H, s), 6.85 (1H, s); .sup.13C-NMR
(75 MHz, CDCl.sub.3) .delta.: 30.02, 31.60, 35.53, 51.08, 56.21,
63.73, 65.35, 79.83, 104.55, 108.37, 110.10, 116.56, 121.85,
128.61, 143.22, 145.65, 145.71; IR (neat): 3536 cm.sup.-1; MS (EI):
m/z 341 (M.sup.+); HRMS (EI): calcd for C.sub.19H.sub.19NO.sub.5
341.1297, found 341.1263.
Example 3
Synthesis of
9-hydroxy-8-methoxy-3-[1,3]dioxolane-2,3,5a,10c-tetrahydro-1H,6H-5-oxa-3--
oxo-acephenanthrylene-10b-carbonitrile ethylene ketal (Compound
22)
##STR00030##
[0153] Under an argon atmosphere, to a THF solution (2.0 mL) of
Compound 41 (41.4 mg, 83.2 .mu.mol), TBAF (1.0 M THF solution, 108
.mu.L, 108 .mu.mol) was added dropwise at 0.degree. C., the whole
was stirred at room temperature for 30 minutes, and then the
reaction was stopped by a saturated aqueous NH.sub.4Cl solution.
The organic phase extracted with CH.sub.2Cl.sub.2 was washed with
H.sub.2O, dried over MgSO.sub.4, and filtered, and then the solvent
was removed by evaporation. The residue was purified by silica gel
column chromatography (AcOEt:hexane=1:1) to produce Compound 22
(30.6 mg, 89.6 .mu.mol, 100%) as colorless oil.
[0154] .sup.1H-NMR (300 MHz, CDCl.sub.3) .delta.: 1.80-1.98 (2H,
m), 2.50 (1H, dt, J=4.4 Hz J=14.6 Hz), 2.80 (1H, td, J=3.3 Hz
J=14.6 Hz), 3.06 (1H, dd, J=2.2 Hz J=15.9 Hz), 3.22 (1H, dd, J=3.3
Hz J=15.9 Hz), 3.88 (3H, s), 3.82-3.94 (4H, m), 3.95-4.13 (1H, m),
5.28 (1H, ddd, J=2.2 Hz J=3.3 Hz J=10.4 Hz), 5.62 (1H, s), 6.04
(1H, d, J=2.2 Hz), 6.76 (1H, s), 6.90 (1H, s); .sup.13C-NMR (75
MHz, CDCl.sub.3) .delta.: 29.74, 31.31, 33.42, 35.26, 50.87, 55.86,
63.59, 65.19, 79.74, 104.48, 110.32, 112.11, 112.40, 121.93,
123.03, 126.85, 142.80, 144.58, 146.25; IR (neat): 3444 cm.sup.-1;
MS (EI): m/z 341 (M.sup.+).
Example 4
Measurement of Neuronal Cell Death Induced by A.beta.(25-35)
[0155] Cerebral cortical neurons of SD rats (embryonic day 18) were
primary cultured. The neurons were seeded in a 24-well culture dish
at a density of 2.86.times.10.sup.5 cells/cm.sup.2, and after 3
days, simultaneously treated with 10 .mu.M partial peptide
including the 25 to 35-amino acid residue of A.beta. (hereinafter
called A.beta.(25-35)) and each of Compounds 1 to 5 (100 nM). After
another 4 days, 6 .mu.M calcein AM was added, the whole was
incubated for 40 minutes at 37.degree. C., and the fluorescence
intensity at 485 nm was measured so that the uptake of calcein AM
into the neurons was calculated to determine the cell viability.
The group without the A.beta.(25-35) treatment and the
administration of any medical agents was defined as a control
group, and the group with the A.beta.(25-35) treatment but without
the administration of any medical agents was defined as a vehicle
group.
[0156] Separately, the neurons were seeded in an 8-well culture
slide at a density of 2.14.times.10.sup.5 cells/cm.sup.2, and after
5 days, simultaneously treated with 10 .mu.M A.beta.(25-35) and
Compound 11 (100 nM). After another 4 days, 6 .mu.M calcein AM was
added, and the whole was incubated for 40 minutes at 37.degree. C.
Then the neurons were fixed, and the fluorescence image was
obtained under a fluorescence microscope AX-80. The uptake of
calcein AM into the neurons was quantitatively analyzed with an
ATTO Densitograph (ATTO, Tokyo, Japan) to determine the cell
viability.
[0157] Here, the data is shown by mean value.+-.standard error. The
significance test was carried out by one way ANOVA (analysis of
variance) and then by Dunnett's test as post hoc test. The
significance level was 5%.
[0158] The results are shown in FIG. 1 and FIG. 2. The vehicle
group was ascertained to have significantly low cell viability. In
contrast, each of the groups treated with Compounds was ascertained
to have higher cell viability comparing with that of the vehicle
group. In particular, the group treated with Compound 11 was
ascertained to have a cell viability remarkably higher than that of
the control group.
Example 5
Measurement of Dendrite Atrophy Induced by A.beta.(25-35)
[0159] Cerebral cortical neurons of SD rats (embryonic day 20) were
primary cultured. The neurons were seeded in an 8-well culture
slide at a density of 1.43.times.10.sup.5 cells/cm.sup.2, and after
3 days, treated with 10 .mu.M A.beta.(25-35) and each of Compounds
5 and 11 (10 .mu.M). After another 5 days, immunostaining was
carried out. The used primary antibody was a rabbit polyclonal
antibody against MAP2 that is a marker protein of the dendrite (a
dilution degree of 1000; Chemicon, Temecula, Calif., USA). The used
secondary antibody was Alexa Fluor 568 labeled goat anti-rabbit IgG
(a dilution degree of 300; Molecular Probes, Carlsbad, Calif.,
USA). The neurons were observed under a fluorescence microscope
AX-80 (Olympus, Tokyo, Japan) and the fluorescence image was
obtained. The average length of the dendrites per neuron was
measured with Neurocyte Ver. 1.5 (Toyobo, Osaka, Japan). The group
without the A.beta.(25-35) treatment and the administration of any
medical agents was defined as a control group, and the group with
the A.beta.(25-35) treatment but without the administration of any
medical agents was defined as a vehicle group.
[0160] The results are shown in FIG. 3. The vehicle group was
ascertained to have the dendrite atrophy. In contrast, each of the
groups treated with Compounds was ascertained to have dendrite
outgrowth effect as remarkably high as that of the control
group.
Example 6
Measurement of Axon Atrophy Induced by A.beta.(25-35)
[0161] Cerebral cortical neurons of SD rats (embryonic day 20) were
primary cultured. The neurons were seeded in an 8-well culture
slide at a density of 2.14.times.10.sup.5 cells/cm.sup.2, and after
3 days, treated with 10 .mu.M A.beta.(25-35) and each of Compounds
8 (1 .mu.M) and 11 (1 and 10 .mu.g/ml). After another 4 days,
immunostaining was carried out. The used primary antibody was a
rabbit polyclonal antibody against phosphorylated neurofilament H
that is a marker protein of the axon (a dilution degree of 300;
Sternberger Monoclonals, MD, USA). The used secondary antibody was
Alexa Fluor 488 labeled goat anti-rabbit IgG (a dilution degree of
300; Molecular Probes, Carlsbad, Calif., USA). The neurons were
observed under a fluorescence microscope AX-8.0 (Olympus, Tokyo,
Japan) and the fluorescence image was obtained. The average length
of the axons per neuron was measured with Neurocyte Ver. 1.5
(Toyobo, Osaka, Japan). The group without the A.beta.(25-35)
treatment and the administration of any medical agents was defined
as a control group, and the group with the A.beta.(25-35) treatment
but without the administration of any medical agents was defined as
a vehicle group.
[0162] The results are shown in FIG. 4. The vehicle group was
ascertained to have the axon atrophy. In contrast, each of the
groups treated with Compounds was ascertained to have axon
outgrowth effect as remarkably high as that of the control
group.
Example 7
Measurement of Dendrite Atrophy Induced by A.beta.(1-42)
[0163] Cerebral cortical neurons of SD rats (embryonic day 18) were
primary cultured. The neurons were seeded in an 8-well culture
slide at a density of 2.6.times.10.sup.5 cells/cm.sup.2. After 5
days, the neurons were treated with 5 .mu.M A.beta.(1-42), and
after another 3 days, treated with each of Compounds 5, 11, 21, and
22 (each 1 .mu.M). After another 5 days, immunostaining was carried
out. The used primary antibody was a rabbit polyclonal antibody
against MAP2 that is as a marker protein of the dendrite (a
dilution degree of 500; Chemicon, Temecula, Calif., USA). The used
secondary antibody was Alexa Fluor 568 labeled goat anti-rabbit IgG
(a dilution degree of 300; Molecular Probes, Carlsbad, Calif.,
USA). The neurons were observed under a fluorescence microscope
AX-80 (Olympus, Tokyo, Japan) and the fluorescence image was
obtained. The average length of the dendrites per neuron was
measured with Neurocyte Ver. 1.5 (Toyobo, Osaka, Japan). The group
without the A.beta.(1-42) treatment and the administration of any
medical agents was defined as a control group, and the group with
the A.beta.(1-42) treatment but without the administration of any
medical agents was defined as a vehicle group.
[0164] The results are shown in FIG. 5. The vehicle group was
ascertained to have the dendrite atrophy. In contrast, each of the
groups treated with Compounds was ascertained to have dendrite
outgrowth effect as remarkably high as that of the control group.
In particular, the group treated with Compound 22 was ascertained
to have a dendrite outgrowth effect remarkably higher than that of
the control group.
Example 8
Measurement of Axon Atrophy Induced by A.beta.(1-42)
[0165] Cerebral cortical neurons of SD rats (embryonic day 18) were
primary cultured. The neurons were seeded in an 8-well culture
slide at a density of 2.6.times.10.sup.5 cells/cm.sup.2, after 5
days, treated with 5 .mu.M A.beta.(1-42), and after another 3 days,
treated with each of Compounds 5, 11, 21, and 22 (each 1 .mu.M).
After another 5 days, immunostaining was carried out. The used
primary antibody was a mouse monoclonal antibody against
phosphorylated neurofilament H that is a marker protein of the axon
(a dilution degree of 500; Sternberger Monoclonals, MD, USA). The
used secondary antibody was Alexa Fluor 488 labeled goat
anti-rabbit IgG (a dilution degree of 300; Molecular Probes,
Carlsbad, Calif., USA). The neurons were observed under a
fluorescence microscope AX-80 (Olympus, Tokyo, Japan) and the
fluorescence image was obtained. The average length of the axons
per neuron was measured with Neurocyte Ver. 1.5 (Toyobo, Osaka,
Japan). The group without the A.beta.(1-42) treatment and the
administration of any medical agents was defined as a control
group, and the group with the A.beta.(1-42) treatment but without
the administration of any medical agents was defined as a vehicle
group.
[0166] The results are shown in FIG. 6. The vehicle group was
ascertained to have the axon atrophy. In contrast, each of the
groups treated with Compounds was ascertained to have axon
outgrowth effect as remarkably high as that of the control group.
In particular, the group treated with Compound 22 was ascertained
to have axon outgrowth effect remarkably higher than that of the
control group.
[0167] The above results reveal that the compound of general
formula (1) has the remarkable neuronal cell death inhibition
effect and the neurite outgrowth effect, and the compound of
general formula (1) is ascertained to be useful for the therapeutic
and/or prophylactic agent for neurological disorders.
* * * * *